Magnetic sorting of membrane associated IgG for phenotype-based selection of stable antibody producing cells

J Immunol Methods. 2017 May:444:1-6. doi: 10.1016/j.jim.2017.02.004. Epub 2017 Feb 9.

Abstract

To establish a simple and widely accessible technique for rapidly selecting high producing Chinese hamster ovary (CHO) cells engineered to express a monoclonal antibody (mAb), we have exploited the transient display of recombinant protein on their cell surface. In combination with magnetic bead-based methods, we demonstrate the ability to select for cells of high productivity in the absence of any metabolic-based selection method. This technique is sufficient to obtain genetically stable engineered CHO cells via a single step of cell subcloning and yields sought-after stable, high IgG producing clonal cell lines. This technique may also be applied to other types of cells as well as polyclonal Ab cell pools.

Keywords: Cell line stability; Magnetic cell sorting; Metabolic selection free expression; Recombinant antibody production.

MeSH terms

  • Animals
  • Antibody-Producing Cells / immunology
  • Antibody-Producing Cells / metabolism*
  • CHO Cells
  • Cell Membrane / immunology
  • Cell Membrane / metabolism*
  • Cloning, Molecular / methods*
  • Cricetulus
  • Genes, Reporter
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / genetics
  • Humans
  • Immunoglobulin G / biosynthesis*
  • Immunoglobulin G / genetics
  • Immunoglobulin G / immunology
  • Immunoglobulin Heavy Chains / biosynthesis*
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Heavy Chains / immunology
  • Immunoglobulin lambda-Chains / biosynthesis*
  • Immunoglobulin lambda-Chains / genetics
  • Immunoglobulin lambda-Chains / immunology
  • Immunomagnetic Separation / methods*
  • Phenotype
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / immunology
  • Transfection

Substances

  • Immunoglobulin G
  • Immunoglobulin Heavy Chains
  • Immunoglobulin lambda-Chains
  • Recombinant Proteins
  • Green Fluorescent Proteins